| Steps 48–62 | Reagents to prepare |
|---|---|
| Coverslip removal | Prewarm 0.2x SSC buffer and 0.2x SSC buffer + 20% deionized formamide at 50°C (step 48) |
| Washes | 0.2x SSC buffer + 20% deionized formamide, 1x PBS and 1x TBS (steps 49-51 and 57) |
| Stop hybridization and detection | Blocking buffer (step 52), anti-DIG buffer (step 54) and washing buffer (steps 53, 55-56) |
| Raising tissue pH | 1x TN (step 58) |
| Development | NBT/BCIP staining solution (step 59) |
| Treatment stop buffer | 1x TE (step 60) |
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